Transformation involving bacteria is a process that inserts a plasmid of DNA into a colony of bacteria with the intention of forcing the bacteria to become competent, and exhibit traits that were coded on the plasmid. Competent bacteria can naturally take in free DNA and gain antibiotic resistance, and even glow. (Sigma-Aldrich 2018). Transformation is used in many applications including the synthesis of essential proteins needed in the human body and to clone DNA needed to continue the process. (SLH 2007). In this experiment, E. coli will undergo the transformation process to insert a plasmid (pGLO) coded with ampicillin resistance, the ability to process arabinose, and the ability to synthesis green fluorescent protein (GFP). Ampicillin is an antibiotic that acts as an inhibitor on the E. coli’s ability …show more content…
The tubes were immediately put in an ice until E. coli was added. Then, we added a single colony of bacteria to both tubes and added the plasmid to the tube labeled +pGLO and returned the tubes to the ice bath for ten minutes. Next, four agar plates were prepared: one that had ampicillin (amp) and arabinose (ara), two with just ampicillin, and one with no alterations. After ten minutes, the bacteria were moved to a hot water bath for 50 seconds and then put immediately back on ice for two minutes. After two minutes, 250 microliters of nutrient broth were added to both tubes and the bacteria incubated at room temperature for ten minutes. After ten minutes, 100 microliters of the +pGLO solution were added to the plate with ampicillin and arabinose and one of the plates with just ampicillin, and 100 microliters of the -pGLO solution were added to the other plate containing ampicillin and the control plate. These plates were left to incubate overnight. (Bio-Rad
Once the recombinant plasmid was obtained, it was then inserted into E. coli cells through transformation. From a successful transformation, we expected the bacterial cells to translate the inserted EGFP sequence into its protein form. The bacteria cultures were plated on petri dishes containing growth supplement, Luria Broth (LB), an antibiotic: Kanamycin, and IPTG which induced the fluorescence property within successfully transformed bacterial colonies. Different variants of the petri dishes were also included as control and unknown.
The plasmids in lanes 3,4,8 and 9 have been digested using one restriction enzyme and had been cut at one restriction site, resulting in a linear molecule. Comparing lanes 3 and 4 to
The first day an unknown sample was assigned to each group of students. The first test applied was a gram stain to test for gram positive or gram-negative bacteria. The morphology of the two types of bacteria was viewed under the microscope and recorded. Then the sample was put on agar plates using the quadrant streak method for isolation. There were three agar plates; one was incubated at room temperature, the second at 30 degrees Celsius, and the third at 37 degrees Celsius. By placing each plate at a different temperature optimal growth temperature can be predicted for both species of bacteria.
The plate with –pGLO, LB and ampicillin will show no growth and no glowing as no arabinose is present for glowing to be activated or ampicillin for growth promotion. Not only to make things glow in the dark when activated, but to enhance crop yield in developing countries and cure illnesses in people who cannot naturally fight them. The main weakness of our experimental procedure was not correctly mixing the plasmid correctly or not effectively to promote genetic transformation, but other weaknesses were prevalent throughout the experiment also.
At the given time sets, CTAB was added to the tubes to kill the E. coli cells and lyse the cells to release its contents including galactosidase enzyme.
Sterile compounding is the preparation of products that should be free from all viable forms of life. There are more stringent requirements for sterile compounding than there are for non-sterile compounding. Staff must be trained and tested on their aseptic processing abilities, cleaner aseptic facilities are required, the quality of air entering the aseptic facility must be evaluated and maintained, sterilisation processes must be effective, knowledge of solution stability is needed and sterility testing of the products is required. The most common type of compounded sterile preparations (CSP) used clinically are aqueous injections. These CSPs require greater attention when being prepared as they pose the greatest risk to the patient if they are non-sterile or contain the wrong ingredients and/or wrong concentrations of ingredients if they are given intravenously. The main objective of sterile compounding is to prevent both morbidity and mortality of patients, which can be caused by non-sterility of preparations, high bacterial endotoxin content and errors associated with ingredients of the preparation, as mentioned earlier.
The purpose of this experiment is to identify an unknown insert DNA by using plasmid DNA as a vector to duplicate the unknown insert DNA. The bacteria will then be transformed by having it take in the plasmid DNA, which will allow us to identify our unknown insert as either the cat gene or the kan gene.
A Comparison of the Laboratory and Industrial Processes When going through the process of fermentation in a laboratory they use certain methods to achieve their goals and some of the methods that they use are completely different from the ones that are used in the industry of fermentation. A fermenter is a container that maintains optimum conditions needed to grow a particular organism I will be using different criteria’s to compare the laboratory and industrial process of fermentation in this assignment; some of them are listed below: * Equipment Used * The Quantity of the Product * Method Used * Quality of the Product Before I get right on into the assignment I will firstly talk about penicillin is and what it is used for today in our society because penicillin will come up. Penicillin was discovered by Alexander Fleming in 1929 and penicillin is one of the earliest discovered and widely used antibiotic agents, derived from the penecillium mold and the use of penecillium did not begin until the 1940s. Penicillin kills bacteria by interfering with the ability to synthesis the cell wall and this will disallow it from splitting and reproducing and it will only lengthen longer Below are is a table that shows the most obvious differences in fermentation in a laboratory and fermentation in the scientific industry: Laboratory Fermentation: Industry Fermentation: It is a batch culture They use a Ph sensor The Ph level is not being controlled The equipment used is more expensive The temperature is not being measured They use a thermometer The yeast population isn’t been given O² They equip the fermenter with an exit gas and an exit liquid flow The food supply is not being replenished They also equip it with a antifoam and gas flow It also has a dissolved O² sensor Equipped with an Sparser In industry they have a fresh media feed
This experiment synthesized luminol (5-Amino-2,3-dihydro-1,4-phthalazinedione) and used the product to observe how chemiluminescence would work. The starting material was 5-nitro-2,3-dihydrophthalazine-1,4-dione, which was, after addition of reaction agents, refluxed and vacuum filtered to retrieve luminol. Using two stock solutions, we missed our precipitated luminol with sodium hydroxide, potassium ferricyanide, and hydrogen peroxide, in their respective solutions, in a dark room, to observe the blue light
Sometimes genes can also be transformed by viruses that can extract a gene from one bacterial cell and inject it into another (3).... ... middle of paper ... ... This understanding must extend to the need to preserve microbial communities that are susceptible to antibiotics, so they will always be able to out-compete resistant strains.
Also, they were able to find the potential of penicillin as a medicine. As a result of Florey and Chain’s research and development of penicillin as a medicine, the production and usage of the drug
The “Fast Plant” experiment is an observation of a plants growth over the span of twenty-eight days. The objective is to observe how plants grow and use their resources throughout the span of their life. In our lab we observed the Brassica rapa, a herbaceous plant in the mustard family which has a short cycle which makes it a perfect plant to observe in this experiment. Like other plants the Brassica rapa must use the resources in the environment to create energy to complete itʻs life cycle and reproduce. By observing the plant it is easy to see in what organ or function the plant is using itʻs energy and resources and if overtime the resources switch to other part of the plants. By conducting this experiment we are able to observe where and how plants allocate their resources throughout their life by harvesting plants at different points in their life.
In modern society, living a long life is meaningless since the quality of life is the axis, which defines a happy life. Consequently, drugs and exercises are being continuously developed and the environment is becoming a vital issue. As our method of life is becoming more complex, unexpected accidents happen and diverse lifestyles are causing people to lose their health. Furthermore, there are incidents where individuals lose some functions of the body or have to live an uncomfortable life due to an infection or a disease. Thus, two experiments were held to solve the question of regeneration, in order to recover body functions and strengthening immunity, in order to fight back diseases more successfully. The first experiment was about regeneration of the skin and the
The birth of genetic engineering and recombinant DNA began in Stanford University, in the year 1970 (Hein). Biochemistry and medicine researchers were pursuing separate research pathways, yet these pathways converged to form what is now known as biotechnology (Hein). The biochemistry department was, at the time, focusing on an animal virus, and found a method of slicing DNA so cleanly that it would reform and go on to infect other cells. (Hein) The medical department focused on bacteria and developed a microscopic molecular messenger, that could not only carry a foreign “blueprint”, or message, but could also get the bacteria to read and copy the information. (Hein) One concept is needed to understand what happened at Stanford: how a bacterial “factory” turns “on” or “off”. (Hein) When a cell is dividing or producing a protein, it uses promoters (“on switches”) to start the process and terminators (“off switches”) to stop the process. (Hein) To form proteins, promoters and terminators are used to tell where the protein begins and where it ends. (Hein) In 1972 Herbert Boyer, a biochemist, provided Stanford with a bacterial enzyme called Eco R1. (Hein) This enzyme is used by bacteria to defend themselves against bacteriophages, or bacterial viruses. (Hein) The biochemistry department used this enzyme as a “molecular scalpel”, to cut a monkey virus called SV40. (Hein) What the Stanford researchers observed was that, when they did this, the virus reformed at the cleaved site in a circular manner. It later went on to infect other cells as if nothing had happened. (Hein) This proved that EcoR1 could cut the bonding sites on two different DNA strands, which could be combined using the “sticky ends” at the sites. (Hein). The contribution towards genetic engineering from the biochemistry department was the observations of EcoR1’s cleavage of
Introduction: A phase change is a result from the kinetic energy (heat) either decreasing or increasing to change the state of matter (i.e. water, liquid, or gas.) Thus saying, freezing is the phase change from a liquid to a solid which results from less kinetic energy/heat. Also, melting is the phase change from a solid to a liquid which results from adding kinetic energy/heat. So, the freezing and melting point of something is the temperature at which these phase changes occur. Therefore, a phase change will occur when a vial of 10 mL of water is placed into a cup of crushed ice mixed with four spoonfuls with 5 mL of sodium chloride for 30 minutes. If 10 mL of water is placed in an ice bath, it will then freeze at 5 degrees Celsius because the kinetic energy will leave quicker with the ice involved. The purpose of this lab is to observe what temperature the water must be to undergo a phase change.